Substituted thiophene carboxylic amide glucagon receptor antagonists, preparation and therapeutic uses

ABSTRACT

The present invention discloses novel compounds of Formula (I), or pharmaceutically acceptable salts thereof, which have glucagon receptor antagonist or inverse agonist activity, as well as methods for preparing such compounds. In another embodiment, the invention discloses pharmaceutical compositions comprising compounds of Formula (I) as well as methods of using them to treat diabetic and other glucagon related metabolic disorders, and the like.

This is the national phase application, under 35 USC 371, forPCT/US2006/061132, filed Nov. 21, 2006, which claims the benefit, under35 USC 119(e), of U.S. provisional application 60/739,692 filed Nov. 23,2005.

This invention relates to compounds that are antagonists or inverseagonists of the glucagon receptor, and to pharmaceutical compositionsthereof, and the uses of these compounds and compositions in thetreatment of the human or animal body. The present compounds show a highaffinity and selective binding for the glucagon receptor, and as suchare useful in the treatment of disorders responsive to the modulation ofglucagon receptors, such as diabetic and other glucagon relatedmetabolic disorders, and the like.

Glucagon is a key hormonal agent that, in cooperation with insulin,mediates homeostatic regulation of the amount of glucose in the blood.Glucagon primarily acts by stimulating certain cells (important amongthese are liver cells) to release glucose when blood glucose levelsfall. The action of glucagon is opposite to that of insulin, whichstimulates cells to take up and store glucose whenever blood glucoselevels rise. Both glucagon and insulin are peptide hormones. Glucagon isproduced in the alpha islet cells of the pancreas and insulin isproduced in the beta islet cells. Glucagon exerts its action by bindingto and activating its receptor, which is a member of theGlucagon-Secretin branch of the 7-transmembrane G-protein coupledreceptor family. The receptor functions by activating the adenylylcyclase second messenger system resulting in an increase in cAMP levels.The glucagon receptor, or naturally occurring variants of the receptor,may possess intrinsic constitutive activity, in vitro, as well as invivo (i.e. activity in the absence of an agonist). Compounds acting asinverse agonists can inhibit this activity. Diabetes mellitus is acommon disorder of glucose metabolism. The disease is characterized byhyperglycemia and may be classified as type 1 diabetes, theinsulin-dependent form, or type 2 diabetes, which isnon-insulin-dependent in character. Subjects with type 1 diabetes arehyperglycemic and hypoinsulinemic, and the conventional treatment forthis form of the disease is to provide insulin. However, in somepatients with type 1 or type 2 diabetes, absolute or relative elevatedglucagon levels have been shown to contribute to the hyperglycemicstate. Both in healthy control animals as well as in animal models oftype 1 and type 2 diabetes, removal of circulating glucagon withselective and specific antibodies has resulted in reduction of theglycemic level. Mice with a homozygous deletion of the glucagon receptorexhibit increased glucose tolerance. Also, inhibition of glucagonreceptor expression using antisense oligonucleotides amelioratesdiabetic syndrome in db/db mice. These studies suggest that glucagonsuppression or an action that antagonizes glucagon could be a usefuladjunct to conventional treatment of hyperglycemia in diabetic patients.The action of glucagon can be suppressed by providing an antagonist oran inverse agonist, i.e. substances that inhibit or preventconstitutive, or glucagon-induced, glucagon receptor mediated responses.

Several publications disclose peptides that are stated to act asglucagon antagonists. Peptide antagonists of peptide hormones are oftenpotent; however they are generally known not to be orally availablebecause of degradation by physiological enzymes and poor distribution invivo. Therefore, orally available non-peptide antagonists of peptidehormones are generally preferred.

A number of publications have appeared in recent years reportingnon-peptide agents that act at the glucagon receptor. For example, WO03/048109, WO 2004/002480, and Kurukulasuriya et al., “Biaryl amideglucagon receptor antagonists” Bioorganic & Medicinal Chemistry Letters,vol. 14, no. 9, pages 2047-2050, 2004, each disclose non-peptidecompounds allegedly having glucagon receptor antagonist activity. Inspite of the number of treatments for diseases that involve glucagon,the current therapies suffer from one or more inadequacies, includingpoor or incomplete efficacy, unacceptable side effects, andcontraindications for certain patient populations. Thus, there remains aneed for an improved treatment using alternative or improvedpharmaceutical agents that modulate glucagon receptor activity and treatthe diseases that could benefit from glucagon receptor modulation. Thepresent invention provides such a contribution to the art based on thefinding that a novel class of compounds has a high affinity, selective,and potent inhibitory activity at the glucagon receptor. The presentinvention is distinct in the particular structures and their activities.

SUMMARY OF THE INVENTION

The present invention provides a compound structurally represented byFormula I:

or a pharmaceutically acceptable salt thereof wherein:R1 and R2 are independently —H or -halogen;R3 is

-   -   —(C₁-C₈) alkyl(optionally substituted with 1 to 3 halogens),        —(C₃-C₇)cycloalkyl,    -   —(C₁-C₆)alkyl-(C₃-C₇)cycloalkyl, or        —(C₃-C₇)cycloalkyl-(C₁-C₆)alkyl(optionally substituted with 1 to        3 halogens);        R4 and R5 are independently    -   —H, -halogen, -hydroxy, hydroxymethyl, —CN, —(C₁-C₇) alkoxy,        —(C₂-C₇)alkenyl, or    -   —(C₁-C₆)alkyl (optionally substituted with 1 to 3 halogens);        R6 is

wherein the zig-zag mark shows the point of attachment to the parentmolecule;R7 and R8 are independently

-   -   —H, -halogen, —(C₁-C₆)alkyl(optionally substituted with 1 to 3        halogens),    -   —(C₁-C₆)alkoxy, —(C₃-C₇)cycloalkyl, —C(O)R10, —COOR10,        —OC(O)R10,    -   —OS(O)₂R10, —SR10, —S(O)R10, —S(O)₂R10, or —O(C₂-C₇)alkenyl;        R9 is independently    -   —H, -halogen, —CN, —(C₃-C₇)cycloalkyl, —C(O)R10, —COOR10,        —OC(O)R10,    -   —OS(O)₂R10, —SR10, —S(O)R10, —S(O)₂R10, or —O(C₂-C₇)alkenyl,    -   —(C₁-C₃)alkoxy(optionally substituted with 1 to 3 halogens), or        —(C₁-C₆)alkyl (optionally substituted with 1 to 3 halogens); and        R10 is independently at each occurrence    -   -hydrogen, or —(C₁-C₆)alkyl(optionally substituted with 1 to 3        halogens).

The present invention provides compounds that are useful as glucagonreceptor antagonists or inverse agonists. The present invention furtherprovides compounds that are selective antagonists or inverse agonists ofthe glucagon receptor over the GLP-1 receptor. The present inventionfurther provides a pharmaceutical composition which comprises a compoundof Formula I, or a pharmaceutical salt thereof, and a pharmaceuticallyacceptable carrier, diluent, or excipient. The present invention furtherprovides methods of using these compounds and compositions in thetreatment of disorders responsive to the modulation of glucagonreceptors, such as diabetic and other glucagon related metabolicdisorders.

DETAILED DESCRIPTION OF THE INVENTION

In one embodiment, the present invention provides compounds of Formula Ias described in detail herein. While all of the compounds of the presentinvention are useful, certain of the compounds are particularlyinteresting and are preferred. The following listing sets out severalgroups of preferred compounds. It will be understood that each of thelistings may be combined with other listings to create additional groupsof preferred embodiments as indicated herein.

In another embodiment the invention provides a compound of formula Iwherein R1 and R2 are —H;

R3 is

-   -   —(C₁-C₈)alkyl(optionally substituted with 1 to 3 halogens),        —(C₃-C₆)cycloalkyl,    -   —(C₁-C₆)alkyl-(C₃-C₆)cycloalkyl, or        —(C₃-C₆)cycloalkyl-(C₁-C₆)alkyl(optionally substituted with 1 to        3 halogens);        R4 and R5 are independently    -   —H, -halogen, or —(C₁-C₆)alkyl (optionally substituted with 1 to        3 halogens);        R6 is

wherein the zig-zag mark shows the point of attachment to the parentmolecule;R7 and R8 are independently

-   -   —H, -halogen, —(C₁-C₃)alkyl(optionally substituted with 1 to 3        halogens), or    -   —(C₁-C₃)alkoxy; and        R9 is independently    -   —H, halogen, or —(C₁-C₆)alkyl (optionally substituted with 1 to        3 halogens).

In another embodiment the invention provides a compound of formula Iwherein R1 and R2 are —H;

R3 is

-   -   —(C₁-C₈)alkyl(optionally substituted with 1 to 3 halogens),        —(C₃-C₆)cycloalkyl,    -   —(C₁-C₆)alkyl-(C₃-C₆)cycloalkyl, or        —(C₃-C₆)cycloalkyl-(C₁-C₆)alkyl(optionally substituted with 1 to        3 halogens);        R4 and R5 are independently    -   —H, -halogen, or —CH₃ (optionally substituted with 1 to 3        halogens);        R6 is

wherein the zig-zag mark shows the point of attachment to the parentmolecule;R7 and R8 are independently —H, or -halogen; andR9 is independently —(C₁-C₆)alkyl (optionally substituted with 1 to 3halogens).

In another embodiment the invention provides a compound of formula Iwherein

R1 and R2 are —H; R3 is —(C₁-C₈)alkyl(optionally substituted with 1 to 3halogens), —(C₃-C₆)cycloalkyl, —(C₁-C₆)alkyl-(C₃-C₆)cycloalkyl, or—(C₃-C₆)cycloalkyl-(C₁-C₆)alkyl(optionally substituted with 1 to 3halogens); R4 and R5 are —CH₃ (optionally substituted with 1 to 3halogens) and each occupies a position adjacent to R6 on the phenyl ringto which R6 is attached;R6 is

wherein the zig-zag mark shows the point of attachment to the parentmolecule;R7 and R8 are —H; and R9 is independently —(C₁-C₆)alkyl (optionallysubstituted with 1 to 3 halogens).

In another embodiment the invention provides a compound of Formula Iwherein R1 and R2 are independently hydrogen or halogen; R3 is methyl,ethyl, propyl, isopropyl, butyl, pentyl, hexyl, heptyl, octyl,3,3-dimethylbutyl, 2-methylpropyl, 3-methyl-butyl, tertbutyl,4-methylpentyl, 2,2-dimethylpropyl, 3,3,3-trifluoropropyl,4,4,4-trifluorbutyl, cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl; R4 and R5 are independently hydrogen, methyl, ethyl,tertbutyl, cyclohexyl, pentyl, isopropoxy, chloro, fluoro, bromo,hydroxy, trifluoromethyl, —CN, methoxy, hydroxymethyl,4-methylpentyloxy, or pentyloxy; R7 and R8 are independently hydrogen,fluoro, chloro, methyl, ethyl, pentyl, isopropyl, tertbutyl,trifluoromethyl, acetyl, 2-methylpropyl, methoxy, cyclohexyl, ortrifluoromethoxy; R9 is hydrogen, bromo, fluoro, methyl, tertbutyl,trifluoromethyl, or isopropyl.

Other embodiments of the invention are provided wherein each of theembodiments described herein above is further narrowed as described inthe following preferences. Specifically, each of the preferences belowis independently combined with each of the embodiments above, and theparticular combination provides another embodiment in which the variableindicated in the preference is narrowed according to the preference.

Preferably R1 is —H. Preferably R1 is fluorine. Preferably R1 ischlorine. Preferably R2 is —H. Preferably R2 is fluorine. Preferably R2is chlorine. Preferably R1 and R2 are —H. Preferably R1 is fluorine andR2 is fluorine.

Preferably R3 is —(C₁-C₈)alkyl(optionally substituted with 1 to 3halogens). Preferably R3 is ethyl, propyl, isopropyl, butyl, tertbutyl,3-methyl-butyl, pentyl, hexyl, heptyl, octyl, 3,3-dimethylbutyl,2-methylpropyl, 4-methylpentyl, 2,2-dimethylpropyl,3,3,3-trifluoropropyl, or 4,4,4-trifluorbutyl. Preferably R3 isisopropyl, butyl, tertbutyl, 3-methyl-butyl, pentyl, 3,3-dimethylbutyl,2-methylpropyl, 4-methylpentyl, 2,2-dimethylpropyl, 3-trifluoropropyl,or 4,4,4-trifluorbutyl. Preferably R3 is isopropyl, 3-methyl-butyl,trifluoropropyl, or 4,4,4-trifluorbutyl.

Preferably R3 is —(C₃-C₇)cycloalkyl. Preferably R3 is cyclopropyl,cyclobutyl, cyclopentyl, or cyclohexyl. Preferably R3 is cyclopropyl.Preferably R3 is cyclobutyl. Preferably R3 is cyclopentyl. Preferably R3is cyclohexyl.

Preferably R3 is —(C₁-C₆)alkyl-(C₃-C₇)cycloalkyl. Preferably R3 is—(C₁-C₃)alkyl-(C₃-C₆)cycloalkyl. Preferably R3 is—(C₁-C₃)alkyl-cyclopropyl. Preferably R3 is —(C₁-C₃)alkyl-cyclobutyl.Preferably R3 is —(C₁-C₃)alkyl-cyclopentyl. Preferably R3 is—(C₁-C₃)alkyl-cyclohexyl.

Preferably R3 is —(C₃-C₇)cycloalkyl-(C₁-C₆)alkyl(optionally substitutedwith 1 to 3 halogens). Preferably R3 is-cyclopropyl-(C₁-C₆)alkyl(optionally substituted with 1 to 3 halogens).Preferably R3 is -cyclobutyl-(C₁-C₆)alkyl(optionally substituted with 1to 3 halogens). Preferably R3 is -cyclopentyl-(C₁-C₆)alkyl(optionallysubstituted with 1 to 3 halogens). Preferably R3 is-cyclohexyl-(C₁-C₆)alkyl(optionally substituted with 1 to 3 halogens).

Preferably R4 is —H, -halogen, -hydroxy, hydroxymethyl, or —(C₁-C₆)alkyl(optionally substituted with 1 to 3 halogens). Preferably R4 is —H,-halogen, or —(C₁-C₃)alkyl (optionally substituted with 1 to 3halogens). Preferably R4 is —H, -halogen, or —CH₃. Preferably R4 is —H.Preferably R4 is fluorine, chlorine, or bromine. Preferably R4 is —CH₃.

Preferably R5 is —H, -halogen, -hydroxy, hydroxymethyl, or —(C₁-C₆)alkyl(optionally substituted with 1 to 3 halogens). Preferably R5 is —H,-halogen, or —(C₁-C₃)alkyl (optionally substituted with 1 to 3halogens). Preferably R5 is —H, -halogen, or —CH₃. Preferably R5 is —H.Preferably R5 is fluorine, chlorine, or bromine. Preferably R5 is —CH₃.

Preferably R4 and R5 are —H. Preferably R4 is halogen and R5 is —H.Preferably R4 is —H and R5 is —CH₃. Preferably R4 and R5 are —CH₃.Preferably R4 and R5 are —CH₃ and each occupies a position adjacent toR6 on the phenyl ring to which R6 is attached.

Preferably R7 is -halogen, —(C₁-C₆)alkyl(optionally substituted with 1to 3 halogens), —(C₁-C₆)alkoxy, —(C₃-C₇)cycloalkyl, —C(O)R10, —COOR10,—OC(O)R10, —OS(O)₂R10, —SR10, —S(O)R10, —S(O)₂R10, or —O(C₂-C₇)alkenyl.Preferably R7 is -halogen, —(C₁-C₆)alkyl(optionally substituted with 1to 3 halogens), or —(C₁-C₆)alkoxy. Preferably R7 is —H or -halogen.Preferably R7 is —H.

Preferably R8 is -halogen, —(C₁-C₆)alkyl(optionally substituted with 1to 3 halogens), —(C₁-C₆)alkoxy, —(C₃-C₇)cycloalkyl, —C(O)R10, —COOR10,—OC(O)R10, —OS(O)₂R10, —SR10, —S(O)R10, —S(O)₂R10, or —O(C₂-C₇)alkenyl.Preferably R8 is -halogen, —(C₁-C₆)alkyl(optionally substituted with 1to 3 halogens), or —(C₁-C₆)alkoxy. Preferably R8 is —H or -halogen.Preferably R8 is —H. Preferably R7 is —H and R8 is —H.

Preferably R9 is —(C₁-C₆)alkyl (optionally substituted with 1 to 3halogens). Preferably R9 is methyl, ethyl, propyl, isopropyl, butyl,tertbutyl, trifluoromethyl, 3-methyl-butyl, pentyl, hexyl,3,3-dimethylbutyl, 2-methylpropyl, 4-methylpentyl, 2,2-dimethylpropyl,3-trifluoropropyl, or 4-trifluorbutyl. Preferably R9 is isopropyl,tertbutyl, or trifluoromethyl.

Preferably R7 is —H, R8 is —H, and R9 is isopropyl, tertbutyl, ortrifluoromethyl.

Preferably R10 is independently at each occurrence—(C₁-C₆)alkyl(optionally substituted with 1 to 3 halogens).

Further embodiments of the invention include the compounds of formulaeX1 to X11. A further embodiment of the invention are any novelintermediate preparations described herein which are useful forpreparing the glucagon receptor antagonists or inverse agonists offormulae I, or X1 to X11.

TABLE 1 Formula Number Structure X1

X2

X3

X4

X5

X6

X7

X8

X9

X10

X11

Due to their interaction with the glucagon receptor, the presentcompounds are useful in the treatment of a wide range of conditions anddisorders in which an interaction with the glucagon receptor isbeneficial. These disorders and conditions are defined herein as“diabetic and other glucagon related metabolic disorders”. One of skillin the art is able to identify “diabetic and other glucagon relatedmetabolic disorders” by the involvement of glucagon receptor mediatedsignaling either in the pathophysiology of the disorder, or in thehomeostatic response to the disorder. Thus, the compounds may find usefor example to prevent, treat, or alleviate, diseases or conditions orassociated symptoms or sequelae, of the endocrinological system, thecentral nervous system, the peripheral nervous system, thecardiovascular system, the pulmonary system, and the gastrointestinalsystem, while reducing and or eliminating one or more of the unwantedside effects associated with the current treatments. “Diabetic and otherglucagon related metabolic disorders” include, but are not limited to,diabetes, type 1 diabetes, type 2 diabetes, hyperglycemia, hyperinsulinemia, beta-cell rest, improved beta-cell function by restoringfirst phase response, prandial hyperglycemia, preventing apoptosis,impaired fasting glucose (IFG), metabolic syndrome, hypoglycemia,hyper-/hypokalemia, normalizing glucagon levels, improved LDL/HDL ratio,reducing snacking, eating disorders, weight loss, polycystic ovariansyndrome (PCOS), obesity as a consequence of diabetes, latent autoimmunediabetes in adults (LADA), insulitis, islet transplantation, pediatricdiabetes, gestational diabetes, diabetic late complications,micro-/macroalbuminuria, nephropathy, retinopathy, neuropathy, diabeticfoot ulcers, reduced intestinal motility due to glucagon administration,short bowel syndrome, antidiarrheic, increasing gastric secretion,decreased blood flow, erectile dysfunction, glaucoma, post surgicalstress, ameliorating organ tissue injury caused by reperfusion of bloodflow after ischemia, ischemic heart damage, heart insufficiency,congestive heart failure, stroke, myocardial infarction, arrythmia,premature death, anti-apoptosis, wound healing, impaired glucosetolerance (IGT), insulin resistance syndromes, syndrome X,hyperlipidemia, dyslipidemia, hypertriglyceridemia,hyperlipoproteinemia, hypercholesterolemia, arteriosclerosis includingatherosclerosis, glucagonomas, acute pancreatitis, cardiovasculardiseases, hypertension, cardiac hypertrophy, gastrointestinal disorders,obesity, diabetes as a consequence of obesity, diabetic dyslipidemia,etc.

In addition, the present invention provides a compound of Formula I, ora pharmaceutical salt thereof, or a pharmaceutical composition whichcomprises a compound of Formula I, or a pharmaceutical salt thereof, anda pharmaceutically acceptable carrier, diluent, or excipient: for use ininhibiting the glucagon receptor; for use in inhibiting a glucagonreceptor mediated cellular response in a mammal; for use in reducing theglycemic level in a mammal; for use in treating a disease arising fromexcessive glucagon; for use in treating diabetic and other glucagonrelated metabolic disorders in a mammal; and for use in treatingdiabetes, obesity, hyperglycemia, atherosclerosis, ischemic heartdisease, stroke, neuropathy, and wound healing. Thus, the methods ofthis invention encompass a prophylactic and therapeutic administrationof a compound of Formula I.

The present invention further provides the use of a compound of FormulaI, or a pharmaceutical salt thereof for the manufacture of a medicamentfor inhibiting the glucagon receptor; for the manufacture of amedicament for inhibiting a glucagon receptor mediated cellular responsein a mammal; for the manufacture of a medicament for reducing theglycemic level in a mammal; for the manufacture of a medicament fortreating a disease arising from excessive glucagon; for the manufactureof a medicament for treating diabetic and other glucagon relatedmetabolic disorders in a mammal; and for the manufacture of a medicamentfor preventing or treating diabetes, obesity, hyperglycemia,atherosclerosis, ischemic heart disease, stroke, neuropathy, andimproper wound healing.

The present invention further provides a method of treating conditionsresulting from excessive glucagon in a mammal; a method of inhibitingthe glucagon receptor in a mammal; a method of inhibiting a glucagonreceptor mediated cellular response in a mammal; a method of reducingthe glycemic level in a mammal; a method of treating diabetic and otherglucagon related metabolic disorders in a mammal; a method of preventingor treating diabetes, obesity, hyperglycemia, atherosclerosis, ischemicheart disease, stroke, neuropathy, and improper wound healing; saidmethods comprising administering to a mammal in need of such treatment aglucagon receptor-inhibiting amount of a compound of Formula I, or apharmaceutically acceptable salt thereof, or a pharmaceuticalcomposition which comprises a compound of Formula I, or a pharmaceuticalsalt thereof, and a pharmaceutically acceptable carrier, diluent, orexcipient.

In addition, the present invention provides a pharmaceutical compositionwhich comprises a compound of Formula I, or a pharmaceutical saltthereof, and a pharmaceutically acceptable carrier, diluent, orexcipient: adapted for use in inhibiting the glucagon receptor; adaptedfor use in inhibiting glucagon receptor mediated cellular responses;adapted for use in reducing the glycemic level in a mammal; adapted foruse in treating diabetic and other glucagon related metabolic disordersin a mammal; and adapted for use in preventing or treating diabetes,obesity, hyperglycemia, atherosclerosis, ischemic heart disease, stroke,neuropathy, and wound healing.

The compound or salt of the present invention further provides adiagnostic agent for identifying patients having a defect in theglucagon receptor, as a therapy to increase gastric acid secretions, andto reverse intestinal hypomobility due to glucagon administration. Theinvention also provides a method for the treatment of disorders ordiseases, wherein a glucagon antagonistic action is beneficial, themethod comprising administering to a subject in need thereof aneffective amount of a compound according to the invention. In anotherembodiment of the invention, the present compounds are used for thepreparation of a medicament for the treatment of any glucagon-mediatedconditions and diseases. In another embodiment of the invention, thepresent compounds are used for the preparation of a medicament for thetreatment of hyperglycemia. In yet another embodiment of the invention,the present compounds are used for the preparation of a medicament forlowering blood glucose in a mammal. The present compounds are effectivein lowering the blood glucose, both in the fasting and the postprandialstage. In still another embodiment of the invention, the presentcompounds are used for the preparation of a pharmaceutical compositionfor the treatment of IGT. In a further embodiment of the invention, thepresent compounds are used for the preparation of a pharmaceuticalcomposition for the treatment of type 2 diabetes. In yet a furtherembodiment of the invention the present compounds are used for thepreparation of a pharmaceutical composition for the delaying orprevention of the progression from IGT to type 2 diabetes. In yetanother embodiment of the invention the present compounds are used forthe preparation of a pharmaceutical composition for the delaying orprevention of the progression from non-insulin requiring type 2 diabetesto insulin requiring type 2 diabetes. In a further embodiment of theinvention the present compounds are used for the preparation of apharmaceutical composition for the treatment of type 1 diabetes. Suchtreatment is normally accompanied by insulin therapy. In yet a furtherembodiment of the invention the present compounds are used for thepreparation of a pharmaceutical composition for the treatment ofobesity. In still a further embodiment of the invention the presentcompounds are used for the preparation of a pharmaceutical compositionfor the treatment of disorders of the lipid metabolism. In still anotherembodiment of the invention the present compounds are used for thepreparation of a pharmaceutical composition for the treatment of anappetite regulation or energy expenditure disorder. In a furtherembodiment of the invention, treatment of a patient with the presentcompounds is combined with diet and/or exercise.

In a further aspect of the invention the present compounds areadministered in combination with one or more further active substancesin any suitable ratios. Such further active substances may for examplebe selected from antidiabetics, antiobesity agents, antihypertensiveagents, agents for the treatment of complications resulting from orassociated with diabetes and agents for the treatment of complicationsand disorders resulting from or associated with obesity. The followinglisting sets out several groups of combinations. It will be understoodthat each of the agents named may be combined with other agents named tocreate additional combinations.

Thus, in a further embodiment of the invention the present compounds maybe administered in combination with one or more antidiabetics.

Suitable antidiabetic agents include insulin, insulin analogues andderivatives such as those disclosed in EP 792 290 (Novo Nordisk A/S),for example N^(εB29)-tetradecanoyl des (B30) human insulin, EP 214 826and EP 705 275 (Novo Nordisk A/S), for example Asp^(B28) human insulin,U.S. Pat. No. 5,504,188 (Eli Lilly), for example Lys^(B28) Pro^(B29)human insulin, EP 368 187 (Aventis), for example Lantus®, which are allincorporated herein by reference, GLP-1 and GLP-1 derivatives such asthose disclosed in WO 98/08871 (Novo Nordisk A/S), which is incorporatedherein by reference, as well as orally active hypoglycemic agents.

The orally active hypoglycemic agents preferably comprise imidazolines,sulphonylureas, biguanides, meglitinides, oxadiazolidinediones,thiazolidinediones, insulin sensitizers, insulin secretagogues, such asglimepiride, α-glucosidase inhibitors, agents acting on theATP-dependent potassium channel of the β-cells for example potassiumchannel openers such as those disclosed in WO 97/26265, WO 99/03861 andWO 00/37474 (Novo Nordisk A/S) which are incorporated herein byreference, or mitiglinide, or a potassium channel blocker, such asBTS-67582, nateglinide, GLP-1 antagonists, DPP-IV (dipeptidylpeptidase-IV) inhibitors, PTPase (protein tyrosine phosphatase)inhibitors, inhibitors of hepatic enzymes involved in stimulation ofgluconeogenesis and/or glycogenolysis, glucose uptake modulators,activators of glucokinase (GK) such as those disclosed in WO 00/58293,WO 01/44216, WO 01/83465, WO 01/83478, WO 01/85706, WO 01/85707, and WO02/08209 (Hoffman-La Roche) or those disclosed in WO 03/00262, WO03/00267 and WO 03/15774 (AstraZeneca), which are incorporated herein byreference, GSK-3 (glycogen synthase kinase-3) inhibitors, compoundsmodifying the lipid metabolism such as antilipidemic agents such as HMGCoA inhibitors (statins), compounds lowering food intake, PPAR(Peroxisome proliferator-activated receptor) ligands including thePPAR-alpha, PPAR-gamma and PPAR-delta substypes, and RXR (retinoid Xreceptor) agonists, such as ALRT-268, LG-1268 or LG-1069.

In another embodiment, the present compounds are administered incombination with insulin or an insulin analogue or derivative, such asN^(εB29)-tetradecanoyl des (B30) human insulin, Asp^(B28) human insulin,Lys^(B28) Pro^(B29) human insulin, Lantus®, or a mix-preparationcomprising one or more of these.

In a further embodiment of the invention the present compounds areadministered in combination with a sulphonylurea such as glibenclamide,glipizide, tolbautamide, chloropamidem, tolazamide, glimepride,glicazide and glyburide.

In another embodiment of the invention the present compounds areadministered in combination with a biguanide, for example, metformin.

In yet another embodiment of the invention the present compounds areadministered in combination with a meglitinide, for example, repaglinideor nateglinide.

In still another embodiment of the invention the present compounds areadministered in combination with a thiazolidinedione insulin sensitizer,for example, troglitazone, ciglitazone, piolitazone, rosiglitazone,isaglitazone, darglitazone, englitazone, CS-011/CI-1037 or T 174 or thecompounds disclosed in WO 97/41097, WO 97/41119, WO 97/41120, WO00/41121 and WO 98/45292 (Dr. Reddy's Research Foundation), which areincorporated herein by reference.

In still another embodiment of the invention the present compounds maybe administered in combination with an insulin sensitizer, for example,such as GI 262570, YM-440, MCC-555, JTT-501, AR-H039242, KRP-297,GW-409544, CRE-16336, AR-H049020, LY510929, MBX-102, CLX-0940, GW-501516or the compounds disclosed in WO 99/19313, WO 00/50414, WO 00/63191, WO00/63192, WO 00/63193 such as ragaglitazar (NN 622 or (−)DRF 2725) (Dr.Reddy's Research Foundation) and WO 00/23425, WO 00/23415, WO 00/23451,WO 00/23445, WO 00/23417, WO 00/23416, WO 00/63153, WO 63196, WO00/63209, WO 00/63190 and WO 00/63189 (Novo Nordisk A/S), which areincorporated herein by reference.

In a further embodiment of the invention the present compounds areadministered in combination with an α-glucosidase inhibitor, forexample, voglibose, emiglitate, miglitol or acarbose.

In another embodiment of the invention the present compounds areadministered in combination with an agent acting on the ATP-dependentpotassium channel of the β-cells, for example, tolbutamide,glibenclamide, glipizide, glicazide, BTS-67582 or repaglinide.

In yet another embodiment of the invention the present compounds may beadministered in combination with nateglinide.

In still another embodiment of the invention the present compounds areadministered in combination with an antilipidemic agent orantihyperlipidemic agent for example cholestyramine, colestipol,clofibrate, gemfibrozil, lovastatin, pravastatin, simvastatin,pitavastatin, rosuvastatin, probucol, dextrothyroxine, fenofibrate oratorvastin.

In still another embodiment of the invention the present compounds areadministered in combination with compounds lowering food intake.

In another embodiment of the invention, the present compounds areadministered in combination with more than one of the above-mentionedcompounds for example in combination with metformin and a sulphonylureasuch as glyburide; a sulphonylurea and acarbose; nateglinide andmetformin; repaglinide and metformin, acarbose and metformin; asulfonylurea, metformin and troglitazone; insulin and a sulfonylurea;insulin and metformin; insulin, metformin and a sulfonylurea; insulinand troglitazone; insulin and lovastatin; etc.

In a further embodiment of the invention the present compounds may beadministered in combination with one or more antiobesity agents orappetite regulating agents.

Such agents may be selected from the group consisting of CART (cocaineamphetamine regulated transcript) agonists, NPY (neuropeptide Y)antagonists, MC4 (melanocortin 4) agonists, MC3 (melanocortin 3)agonists, orexin antagonists, TNF (tumor necrosis factor) agonists, CRF(corticotropin releasing factor) agonists, CRF BP (corticotropinreleasing factor binding protein) antagonists, urocortin agonists, β3adrenergic agonists such as CL-316243, AJ-9677, GW-0604, LY362884,LY377267 or AZ-40140 MSH (melanocyte-stimulating hormone) agonists, MCH(melanocyte-concentrating hormone) antagonists, CCK (cholecystokinin)agonists, serotonin re-uptake inhibitors such as fluoxetine, seroxat orcitalopram, serotonin and noradrenaline re-uptake inhibitors, mixedserotonin and noradrenergic compounds, 5HT (serotonin) agonists,bombesin agonists, galanin antagonists, growth hormone, growth factorssuch as prolactin or placental lactogen, growth hormone releasingcompounds, TRH (thyreotropin releasing hormone) agonists, UCP 2 or 3(uncoupling protein 2 or 3) modulators, leptin agonists, DA agonists(bromocriptin, doprexin), lipase/amylase inhibitors, PPAR (peroxisomeproliferator-activated receptor) modulators, RXR (retinoid X receptor)modulators, TR β agonists, AGRP (Agouti related protein) inhibitors, H3histamine antagonists, opioid antagonists (such as naltrexone),exendin-4, GLP-1 and ciliary neurotrophic factor (such as axokine),cannaboid receptor antagonist for example CB-1 (such as rimonabant). Inanother embodiment the antiobesity agent is dexamphetamine oramphetamine. In another embodiment the antiobesity agent is leptin. Inanother embodiment the antiobesity agent is fenfluramine orexfenfluramine. In still another embodiment the antiobesity agent issibutramine. In a further embodiment the antiobesity agent is orlistat.In another embodiment the antiobesity agent is mazindol or phentermine.In still another embodiment the antiobesity agent is phendimetrazine,diethylpropion, fluoxetine, bupropion, topiramate or ecopipam.

Furthermore, the present compounds may be administered in combinationwith one or more antihypertensive agents. Examples of antihypertensiveagents are β-blockers such as alprenolol, atenolol, timolol, pindolol,propranolol and metoprolol, SCE (angiotensin converting enzyme)inhibitors such as benazepril, captopril, enalapril, fosinopril,lisinopril, quinapril and ramipril, calcium channel blockers such asnifedipine, felodipine, nicardipine, isradipine, nimodipine, diltiazemand verapamil, and α-blockers such as doxazosin, urapidil, prazosin andterazosin. Further reference can be made to Remington: The Science andPractice of Pharmacy, 19^(th) Edition, Gennaro, Ed., Mack PublishingCo., Easton, Pa., 1995.

The compounds of the present invention may be administered incombination with FAS inhibitors.

The compounds of the present invention may also be administered incombination with chemical uncouplers, hormone sensitive lipaseinhibitor, imidazolines, 11-β-hydroxysteroid dehydrogenase inhibitors,lipoprotein lipase activator, AMPK activators, immunosuppresive drugs,nicotinamide, ASIS, anti-androgens or carboxypeptidase inhibitors.

It should be understood that any suitable combination of the compoundsaccording to the invention with diet and/or exercise, one or more of theabove-mentioned compounds are considered to be within the scope of thepresent invention.

General terms used in the description of compounds, compositions, andmethods herein described, bear their usual meanings. Throughout theinstant application, the following terms have the indicated meanings:

“GLP-1” means glucagon-like peptide 1. The term “glucagon receptor”means one or more receptors that interact specifically with glucagon toresult in a biological signal. The term “GLP-1 receptor” means one ormore receptors that interact specifically with glucagon-like peptide 1to result in a biological signal.

The term “glucagon receptor antagonist” means a compound of the presentinvention with the ability to block cAMP production in responseglucagon. The term “glucagon receptor inverse agonist” means a compoundof the present invention with the ability to inhibit the constitutiveactivity of glucagon receptor. The term “selective” antagonist orinverse agonist means a compound having greater affinity for theglucagon receptor as compared to the affinity for the GLP-1 receptor.

In the general formulae of the present document, the general chemicalterms have their usual meanings. For example;

“Halogen” or “halo” means fluoro, chloro, bromo and iodo.

The term “alkyl,” unless otherwise indicated, refers to those alkylgroups of a designated number of carbon atoms of either a straight orbranched saturated configuration. As used herein, “(C₁-C₃)alkyl” are oneto three carbon atoms, such as methyl, ethyl, propyl, n-propyl,isopropyl, and the like and branched or isomeric forms thereof, andoptionally may be substituted with one to three halogens or a designatednumber of substituents as set forth in the embodiments recited herein.“(C₁-C₆)alkyl” are one to six carbon atoms such as methyl, ethyl,propyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl andtert-butyl, pentyl, isopentyl, hexyl, and the like, and branched orisomeric forms thereof, and optionally may be substituted with one tothree halogens or a designated number of substituents as set forth inthe embodiments recited herein. “(C₁-C₈) alkyl” are one to eight carbonatoms, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl,octyl, and the like, and branched or isomeric forms thereof, andoptionally may be substituted with one to three halogens as set forth inthe embodiments recited herein.

The term “(C₃-C₇) cycloalkyl” refers to a saturated or partiallysaturated carbocycle containing one or more rings of from 3 to 7 carbonatoms. Examples of (C₃-C₇) cycloalkyl include but are not limited tocyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl. Theterm “(C₃-C₆) cycloalkyl” refers to a saturated carbocycle ring of from3 to 6 carbon atoms. Examples of (C₃-C₆) cycloalkyl include but are notlimited to cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

The term “(C₁-C₃) alkoxy” represents an alkyl group of one to threecarbon atoms attached through an oxygen bridge, such as methoxy, ethoxy,propoxy, and the like. The term “(C₁-C₆) alkoxy” represents an alkylgroup of one to six carbon atoms attached through an oxygen bridge, suchas methoxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy, pentoxy,and the like. The term “(C₁-C₇) alkoxy” represents an alkyl group of oneto seven carbon atoms attached through an oxygen bridge, such asmethoxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy, pentoxy, andthe like, and may be optionally substituted with three halogens as setforth in the embodiments recited herein.

The term “(C₂-C₇) alkenyl” means hydrocarbon chain of two to sevencarbon atoms of either a straight or branched configuration having atleast one carbon-carbon double bond which may occur at any point alongthe chain, such as ethenyl, propenyl, butenyl, pentenyl, vinyl, alkyl,2-butenyl and the like, and may be optionally substituted with one tothree halogens as set forth in the embodiments recited herein.

The term “optionally substituted,” or “optional substituents,” as usedherein, means that the groups in question are either unsubstituted orsubstituted with one or more of the substituents specified. When thegroups in question are substituted with more than one substituent, thesubstituents may be the same or different. Furthermore, when using theterms “independently,” “independently are,” and “independently selectedfrom” mean that the groups in question may be the same or different.Certain of the herein defined terms may occur more than once in thestructural formulae, and upon such occurrence each term shall be definedindependently of the other.

The term “patient” includes human and non-human animals such ascompanion animals (dogs and cats and the like) and livestock animals.Livestock animals are animals raised for food production. Ruminants or“cud-chewing” animals such as cows, bulls, heifers, steers, sheep,buffalo, bison, goats and antelopes are examples of livestock. Otherexamples of livestock include pigs and avians (poultry) such aschickens, ducks, turkeys and geese. Yet other examples of livestockinclude fish, shellfish and crustaceans raised in aquaculture. Alsoincluded are exotic animals used in food production such as alligators,water buffalo and ratites (e.g., emu, rheas or ostriches). The patientto be treated is preferably a mammal, in particular a human being.

The term “a glucagon receptor mediated cellular response” includesvarious responses by mammalian cells to glucagon stimulation or glucagonreceptor activity. For example “glucagon receptor mediated cellularresponses,” include but are not limited to, release of glucose fromliver, or other cells, in response to glucagon stimulation or glucagonreceptor activity. One of ordinary skill in the art can readily identifyother cellular responses mediated by glucagon receptor activity, forexample by observing a change in the responsive cellular endpoint aftercontacting the cell with an effective dose of glucagon.

The terms “treatment”, “treating” and “treat”, as used herein, includetheir generally accepted meanings, i.e., the management and care of apatient for the purpose of preventing, prohibiting, restraining,alleviating, ameliorating, slowing, stopping, delaying, or reversing theprogression or severity of a disease, disorder, or pathologicalcondition, described herein, including the alleviation or relief ofsymptoms or complications, or the cure or elimination of the disease,disorder, or condition.

“Composition” means a pharmaceutical composition and is intended toencompass a pharmaceutical product comprising the active ingredient(s)including compound(s) of Formula I, and the inert ingredient(s) thatmake up the carrier. Accordingly, the pharmaceutical compositions of thepresent invention encompass any composition made by admixing a compoundof the present invention and a pharmaceutically acceptable carrier.

The term “suitable solvent” refers to any solvent, or mixture ofsolvents, inert to the ongoing reaction that sufficiently solubilizesthe reactants to afford a medium within which to effect the desiredreaction.

The term “unit dosage form” means physically discrete units suitable asunitary dosages for human subjects and other non-human animals, eachunit containing a predetermined quantity of active material calculatedto produce the desired therapeutic effect, in association with asuitable pharmaceutical carrier.

The compounds of the present invention may be chiral, and it is intendedthat any enantiomers, whether pure, partially purified, or racemicmixtures, are included within the scope of the invention. Furthermore,when a double bond or a fully or partially saturated ring system or morethan one center of asymmetry or a bond with restricted rotatability ispresent in the molecule diastereomers may be formed. It is intended thatany diastereomers, as separated, pure or partially purifieddiastereomers or mixtures thereof are included within the scope of theinvention. Furthermore, some of the compounds of the present inventionmay exist in different tautomeric forms and it is intended that anytautomeric forms, which the compounds are able to form, are includedwithin the scope of the present invention. The invention also includestautomers, enantiomers and other stereoisomers of the compounds ofFormula I. Such variations are contemplated to be within the scope ofthe invention.

The compounds of Formula I, when existing as a diastereomeric mixture,may be separated into diastereomeric pairs of enantiomers by, forexample, fractional crystallization from a suitable solvent, for examplemethanol or ethyl acetate or a mixture thereof. The pair of enantiomersthus obtained may be separated into individual stereoisomers byconventional means, for example by the use of an optically active acidas a resolving agent. Alternatively, any enantiomer of a compound ofFormula I may be obtained by stereospecific synthesis using opticallypure starting materials or reagents of known configuration or throughenantioselective synthesis.

The term “enantiomeric enrichment” as used herein refers to the increasein the amount of one enantiomer as compared to the other. A convenientmethod of expressing the enantiomeric enrichment achieved is the conceptof enantiomeric excess, or “ee,” which is found using the followingequation:

${ee} = {\frac{E^{1} - E^{2}}{E^{1} + E^{2}} \times 100}$

wherein E¹ is the amount of the first enantiomer and E² is the amount ofthe second enantiomer. Thus, if the initial ratio of the two enantiomersis 50:50, such as is present in a racemic mixture, and an enantiomericenrichment sufficient to produce a final ratio of 70:30 is achieved, theee with respect to the first enantiomer is 40%. However, if the finalratio is 90:10, the ee with respect to the first enantiomer is 80%. Anee of greater than 90% is preferred, an ee of greater than 95% is mostpreferred and an ee of greater than 99% is most especially preferred.Enantiomeric enrichment is readily determined by one of ordinary skillin the art using standard techniques and procedures, such as gas or highperformance liquid chromatography with a chiral column. Choice of theappropriate chiral column, eluent and conditions necessary to effectseparation of the enantiomeric pair is well within the knowledge of oneof ordinary skill in the art. In addition, the specific stereoisomersand enantiomers of compounds of Formula I, can be prepared by one ofordinary skill in the art utilizing well known techniques and processes,such as those disclosed by J. Jacques, et al., “Enantiomers, Racemates,and Resolutions,” John Wiley and Sons, Inc., 1981, and E. L. Eliel andS. H. Wilen,“ Stereochemistry of Organic Compounds,” (Wiley-Interscience1994), and European Patent Application No. EP-A-838448, published Apr.29, 1998. Examples of resolutions include recrystallization techniquesor chiral chromatography. Unless otherwise indicated, a compoundindicated to be “isomer 1” will be the first isomer eluted from thechiral separation column and “isomer 2” will be the second.

In general, the term “pharmaceutical” when used as an adjective meanssubstantially non-toxic to living organisms. For example, the term“pharmaceutical salt” as used herein, refers to salts of the compoundsof Formula I, which are substantially non-toxic to living organisms. Thepresent invention also encompasses pharmaceutically acceptable salts ofthe present compounds. Pharmaceutically acceptable salts and commonmethodology for preparing them are well known in the art. See e.g., P.Stahl, et al., “Handbook Of Pharmaceutical Salts: Properties, Selection,and Use,” (VCHA/Wiley-VCH, 2002); Berge, S. M, Bighley, L. D., andMonkhouse, D. C., “Pharmaceutical Salts,” J. Pharm. Sci., 66:1, 1977.

The invention also encompasses prodrugs of the present compounds, whichon administration undergo chemical conversion by metabolic processesbefore becoming pharmacologically active substances. In general, suchprodrugs will be functional derivatives of present compounds, which arereadily convertible in vivo into a compound of the present invention.Conventional procedures for the selection and preparation of suitableprodrug derivatives are described, for example in “Design of Prodrugs”,ed. H. Bundgaard, Elsevier, 1985.

The compounds of Formula I, can be prepared by one of ordinary skill inthe art following a variety of procedures, some of which are illustratedin the procedures and schemes set forth below. The particular order ofsteps required to produce the compounds of Formula I is dependent uponthe particular compound to being synthesized, the starting compound, andthe relative liability of the substituted moieties. The reagents orstarting materials are readily available to one of skill in the art, andto the extent not commercially available, are readily synthesized by oneof ordinary skill in the art following standard procedures commonlyemployed in the art, along with the various procedures and schemes setforth below.

The following Schemes, Preparations, Examples and Procedures areprovided to better elucidate the practice of the present invention andshould not be interpreted in any way as to limit the scope of the same.Those skilled in the art will recognize that various modifications maybe made while not departing from the spirit and scope of the invention.All publications mentioned in the specification are indicative of thelevel of those skilled in the art to which this invention pertains.

The optimal time for performing the reactions of the Schemes,Preparations, Examples and Procedures can be determined by monitoringthe progress of the reaction via conventional chromatographictechniques. Furthermore, it is preferred to conduct the reactions of theinvention under an inert atmosphere, such as, for example, argon, or,particularly, nitrogen. Choice of solvent is generally not critical solong as the solvent employed is inert to the ongoing reaction andsufficiently solubilizes the reactants to effect the desired reaction.The compounds are preferably isolated and purified before their use insubsequent reactions. Some compounds may crystallize out of the reactionsolution during their formation and then collected by filtration, or thereaction solvent may be removed by extraction, evaporation, ordecantation. The intermediates and final products of Formula I may befurther purified, if desired by common techniques such asrecrystallization or chromatography over solid supports such as silicagel or alumina.

The skilled artisan will appreciate that not all substituents arecompatible with all reaction conditions. These compounds may beprotected or modified at a convenient point in the synthesis by methodswell known in the art.

The terms and abbreviations used in the instant Schemes, Preparations,Examples and Procedures have their normal meanings unless otherwisedesignated. For example, as used herein, the following terms have themeanings indicated: “psi” refers to pounds per square inch; “min” refersto minutes; “h” or “hr” refers to hours; “TLC” refers to thin layerchromatography; “HPLC” refers to high performance liquid chromatography;“R_(f)” refers to retention factor; “R_(t)” refers to retention time;“δ” refers to part per million down-field from tetramethylsilane; “MS”refers to mass spectrometry; “MS (ES)” refers to electron spray massspectrometry, “UV” refers to ultraviolet spectrometry; “¹H NMR” refersto proton nuclear magnetic resonance spectrometry. In addition; “RT”refers to room temperature; “DEAD” refers to diethylazodicrboxylate;“PPh₃” refers to triphenylphosphine; “ADDP” refers to1,1′-(azodicarbonyl)dipiperidine; “PBu₃” refers to tributylphosphine;“OTF” refers to triflate; “LAH” refers to lithium aluminum hydride;“DIBAL-H” refers to diisobutylaluminum hydride; “KOtBu” refers topotassium t-butoxide; “THF” refers to tetrahydrofuran; “TBP” refers totributylphosphine; “EDCI” refers to1-(3-dimethylaminopropyl)-3-ethylcarbodiamide hydrochloride; “DMAP”refers to dimethylaminopyridine; “HNMe(OMe)” refers toN,N,dimethylhydroxyamine; “CDMT” refers to2-chloro-4,6-dimethoxy-[1,3,5] triazine; “NMM” refers to N-methylmorpholine; “DCM” refers to dichloromethane; “DMSO” refers todimethylsulfoxide; “ET₃N” refers to triethylamine; “DMF” refers todimethylformamide; “PBr₃” refers to phosphorus tribromide; “Et” in aformula refers to ethyl, for example Et₂O refers to diethylether, andEtOAc refers to ethylacetate; “PyBOP” refers tobromo-tris-pyrrolidino-phosphonium hexafluorophosphate; “Me” refers tomethyl as in MeOH which is methanol; “Pd/C” refers to 10% palladium oncarbon. Unless otherwise indicated, isomer 1 refers to the first isomerto be eluted in a chiral separation and isomer 2 refers to the secondisomer to be eluted in a chiral separation.

General Schemes

All of the compounds of the present invention can be chemicallyprepared, for example, by following the synthetic routes set forth inthe Schemes and/or the Preparations and Examples below. However, thefollowing discussion is not intended to be limiting to the scope of thepresent invention in any way. For example, the specific synthetic stepsfor each of the routes described may be combined in different ways, orin conjunction with steps from different schemes, to prepare additionalcompounds of Formula I.

In Scheme I, Step A, a 4-halophenol of formula (1), (X═I or Br) iscoupled with a phenyl boronic acid of formula (2), using a Suzukireaction to provide a biphenyl hydroxy of formula (3). It will berecognized by one skilled in the art that such Suzuki couplings usingaryl halides and phenyl boronic acids can be effected using a widevariety of reaction conditions. Preferred conditions useoxydi-2,1-phenylene)bis(diphenylphosphine) in the presence of palladiumacetate and potassium fluoride, in an inert solvent, such astetrahydrofuran. The reaction is heated at a temperature of 50° C. tothe reflux temperature of the solvent for about 4 to 48 hours undernitrogen.

Alternatively the Suzuki reaction may be effected usingtetrakis(triphenylphosphine)palladium with potassium fluoride undernitrogen. The reaction proceeds in an inert solvent such as toluene orbenzene and water at a temperature of 40° C. to the reflux temperatureof the reaction for about 4 to 48 hours

In Scheme I, Step B, a biphenyl hydroxy of formula (3) is coupled withdimethylthiocarbamoyl chloride, undergoes a thermal rearrangement andsubsequent solvolysis of the dimethyl-thiocarbamic acid esterintermediate to provide a biphenyl thiol of formula (4). The couplingreaction to give the thiocarbamate is accomplished in the presence of4-dimethylaminopyridine with an organic amine such as triethylamine ordiisopropylethylamine. The reaction is performed in an inert solventsuch as dioxane, tetrahydrofuran, benzene or toluene, with dioxane beingpreferred at temperature of 65° C. to the reflux temperature of thesolvent. The resulting thiocarbamate is thermally rearranged intetradecane at a temperature of 200 to 250° C. with the preferredtemperature being about 245° C. to give a dimethyl-thiocarbamic acidester. Solvolysis with sodium methoxide in methanol or sodium ethoxidein ethanol gives the biphenyl thiol of formula (4).

In Scheme II, Step A, an ethyl thiophene 2-carboxylate of formula (5) isalkylated with an aldehyde (R3CHO) to give a secondary alcohol offormula (6). An ethyl thiophene 2-carboxylate of formula (5) is treatedwith lithium diisopropylamide at a temperature of −80 to −70° C. andthen treated with an aldhyde in an inert solvent such astetrahydrofuran. The reaction is allowed to warm to ambient temperatureover about 12 to 24 hours and the product is isolated using extractivetechniques to give the secondary alcohol of formula (6).

In Scheme II, Step B, a secondary alcohol of formula (6) is coupled witha 4-halo thiophenol (X═Br or I) of formula (7), to give a 4-halophenylthioether of formula (8). The coupling is effected with a Lewis acid,such as zinc iodide (ZnI₂), in an inert solvent such as dichloromethaneor dichloroethane at a temperature of 0 to 50° C., with dichloroethaneat room temperature being the preferred conditions. The product isisolated using common extractive techniques to give the 4-halophenylthioether of formula (8).

Alternatively the coupling of the secondary alcohol and thiophenol maybe affected using Mitsinobu conditions. Common redox systems, known tothose skilled in the art, such as diethyl azodicarboxylate(DEAD)/triphenylphospine, N,N,N′,N′-tetramethylazodicarboxamide(TMAD)/tributylphosphine or 1,1′-(azodicarbonyl)dipiperidine(ADDP)/tributylphosphine may be used to effect the transformation.

In Scheme II, Step C, a 4-halophenyl thioether of formula (8), iscoupled with a phenyl boronic acid of formula (2) in a Suzuki reactionto provide the biphenyl thioether of formula (9), using conditions asdescribed for Scheme I, Step A.

Alternatively, in Scheme II, Step D, a biphenyl thiol of formula (3) iscoupled using conditions described for Scheme II, Step B to give abiphenyl thioether of formula (9).

In Scheme II, Step E, the thiophene carboxylic acid ethyl ester offormula (9) is hydrolyzed to a thiophene carboxylic acid of formula(10). The ester is hydrolyzed in an appropriate water soluable solventsuch as ethanol, methanol, dioxane, or tetrahydrofuran, with ethanolbeing preferred. The ester is treated with an inorganic base such aspotassium or sodium hydroxide, with sodium hydroxide being preferred, atroom temperature to the reflux temperature of the solvent for 2 to 48hours. The thiophene carboxylic acid of formula (10) is isolated byneutralization with hydrochloric acid followed by common extractivetechniques.

In Scheme III, Step A, a thiophene carboxylic acid of formula (11) isacylated to give an amide of formula (12). It will be recognized by oneskilled in the art that there are numerous conditions for amide bondformation between a carboxylic acid and an amine. Such methods can befound in the text of R. C. Larock in “Comprehensive OrganicTransformations”, VCH Publishers, 1989, p. 972-976. The preferredconditions use a catalytic amount of 4-dimethylaminopyridine (DMAP),1,[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (EDCI) andan organic base such as diisopropylethylamine or triethylamine in aninert solvent such as dichloromethane or tetrahydrofuran. The activeester is treated with aminoacetonitrile hydrochloride at 0° C. to thereflux temperature of the solvent, but preferably at room temperature,for about 4 to 48 hours.

Alternatively, In Scheme III, Step A, another set of preferredconditions use 2-chloro-4,6-dimethoxy-1,3,5-triazine to form the activeester in the presence of an organic base such as N-methyl morpholine inan inert solvent such as tetrahydrofuran. The active ester is treatedwith aminoacetonitrile hydrochloride at 0 to 50° C. for 4 to 48 hours toform the amide of formula (12).

In Scheme III, Step B, an amide of formula (12) is cyclized to atetrazole of formula (13). It will be recognized by the skilled artisanthat useful reagents for forming tetrazoles from nitriles includeazidotrimethylsilane, azidotributyltin, and sodium azide. The preferredconditions use sodium azide in the presence of an alkyl aminehydrochloride such as triethylamine or diisopropylethylaminehydrochloride in an inert solvent such as toluene, benzene,dimethylformamide, tetrahydrofuran, or dioxane. The preferred conditionsuse toluene at a temperature of 40° C. to the reflux temperature of thesolvent for a period of 4 to 48 hours. The product is isolated byacidification with aqueous hydrochloric acid and extraction into anappropriate organic solvent, such as ethyl acetate.

PREPARATIONS AND EXAMPLES

The Examples provided herein are illustrative of the invention claimedherein and are not intended to limit the scope of the claimed inventionin any way. Names of the preparations and examples are derived usingChemDraw.

¹H NMR spectra are recorded on a Varian 400 MHz spectrometer at ambienttemperature. Data are reported as follows: chemical shift in ppm frominternal standard tetramethylsilane on the (scale, multiplicity(b=broad, s=singlet, d=doublet, t=triplet, q=quartet, qn=quintet andm=multiplet), integration, coupling constant (Hz) and assignment. ¹H-NMRindicates a satisfactory NMR spectrum was obtained for the compounddescribed. Monoisotopic mass spectral data are obtained on an AgilentG1956B MSD single quadrapole instrument using electrospray ionization(ESI or ES). Analytical thin layer chromatography is performed on EMReagent 0.25 mm silica gel 60-F plates. Visualization is accomplishedwith UV light. All examples are racemic unless indicated otherwise.

Preparation 1 2,6-dim ethyl-4′-(trifluoromethyl)biphenyl-4-ol

4-Bromo-3,5-dimethylphenol (115.00 g, 571.96 mmol),4-(trifluoromethyl)phenyl boronic acid (130.36 g, 686.35 mmol),(oxydi-2,1-phenylene)bis(diphenylphosphine) (126.00 g, 233.96 mmol),potassium fluoride (99.69 g, 1.72 mol), and Pd(OAc)₂ (25.68 g, 114.39mmol) are added to nitrogen-sparged tetrahydrofuran (3.0 L) and heatedto reflux. The consumption of the starting material,4-bromo-3,5-dimethylphenol, is monitored by GC. Reflux is maintaineduntil 4-bromo-3,5-dimethylphenol has been consumed and is generallycomplete after 18 h. After the reaction is complete, the batch is cooledto approximately 25° C. The crude reaction mixture is absorbed ontosilica (˜500 g) and eluted over silica (1.5 kg) with 10% ethyl acetatein heptane to obtain the product as a solid (132.9 g, 87.3%). Theproduct is crystallized from heptane (23 L/kg) and isopropanol (0.4L/kg) to yield the title compound (119.5 g; 78.5% yield) as an off-whitesolid. MS (ES): 265.21 [M−1]⁻. ¹H NMR (400 MHz, CDCl₃): δ 7.68 (d, 2H),7.26 (d, 2H), 6.62 (s, 2H), 4.73 (s, 1H), 1.97 (s, 6H).

Preparation 2 4′-tert-butyl-2,6-dimethylbiphenyl-4-ol

Prepare the title compound by essentially following the procedure asdescribed in Preparation 1, using 4-tert-butylphenylboronic acid. ¹H NMR(400 MHz, CDCl₃): δ 7.43 (d, 2H), 7.06 (d, 2H), 6.61 (s, 2H), 4.85 (s,1H), 2.02 (s, 6H), 1.38 (s, 9H).

Preparation 3 (R,S)-5-(1-Hydroxy-3-methyl-butyl)-thiophene-2-carboxylicacid ethyl ester

A solution of diisopropylamine (8.55 mL, 60 mmol) in THF (350 mL) underN₂ is cooled to −78° C. and treated with n-butyllithium (2.5 M inhexanes, 24 mL). The mixture is then warmed to 0° C. for 10 min, cooledback to −78° C., treated dropwise with a solution ofthiophene-2-carboxylic acid ethyl ester (7.8 g, 50 mmol) in THF (150mL), and stirred 5 min. 3-Methyl-butyraldehyde (6.48 mL, 60 mmol) isthen added, and the reaction is allowed to warm to room temperature,while stirring overnight. Aqueous buffer (pH=7) is added, and theproduct is extracted into ethyl acetate (3×). Combined organic layersare dried, filtered, and concentrated. The resulting residue is appliedto silica gel and eluted using hexanes with an ethyl acetate gradientfrom 0% to 60% to give the title compound (8.03 g).

Preparations 4 to 9 are prepared in a substantially similar manner asdescribed in Preparation 3.

Preparation 4 (R,S)-5-(1-Hydroxy-propyl)-thiophene-2-carboxylic acidethyl ester Preparation 5(R,S)-5-(1-Hydroxy-butyl)-thiophene-2-carboxylic acid ethyl esterPreparation 6 (R,S)-5-(1-Hydroxy-pentyl)-thiophene-2-carboxylic acidethyl ester Preparation 7(R,S)-5-(1-Hydroxy-2,2-dimethyl-propyl)-thiophene-2-carboxylic acidethyl ester Preparation 8(R,S)-5-(1-Hydroxy-2-methyl-propyl)-thiophene-2-carboxylic acid ethylester Preparation 9(R,S)-5-(1-Hydroxy-3,3-dimethyl-butyl)-thiophene-2-carboxylic acid ethylester Preparation 10 4′-tert-Butyl-2,6-dimethyl-biphenyl-4-thiol Step A.Dimethyl-thiocarbamic acid O-(4′-tert-butyl-2,6-dimethyl-biphenyl-4-yl)ester

To a solution of 4′-tert-butyl-2,6-dimethyl-biphenyl-4-ol (10 g, 37.3mmol) in dioxane (157 mL) is added 4-dimethylaminopyridine (476 mg, 3.9mmol), triethylamine (10 mL, 78.6 mmol), and dimethylthiocarbamoylchloride (6.1 g, 49.1 mmol). The reaction mixture is heated to refluxovernight. After cooling to room temperature, the reaction mixture ispartitioned between ethyl acetate and water. The aqueous layer isback-extracted with ethyl acetate, and the combined organic layers aredried and concentrated. The resulting residue is applied to silica geland eluted using 20% ethyl acetate in hexanes to givedimethyl-thiocarbamic acid O-(4′-tert-butyl-2,6-dimethyl-biphenyl-4-yl)ester (12.2 g).

Step B. Dimethyl-thiocarbamic acidS-(4′-tert-butyl-2,6-dimethyl-biphenyl-4-yl) ester

A suspension of dimethyl-thiocarbamic acidO-(4′-tert-butyl-2,6-dimethyl-biphenyl-4-yl) ester (12.1 g, 35.4 mmol)in tetradecane (80 mL) is heated at 245° C. for 16 h. After cooling toroom temperature, a solid precipitate is filtered, washed with heptane,and dried under vacuum at 40° C. The resulting residue is applied tosilica gel and eluted using hexanes with an ethyl acetate gradient from0% to 60% to give dimethyl-thiocarbamic acidS-(4′-tert-butyl-2,6-dimethyl-biphenyl-4-yl) ester (8.86 g).

Step C. 4′-tert-Butyl-2,6-dimethyl-biphenyl-4-thiol

To a solution of dimethyl-thiocarbamic acidS-(4′-tert-butyl-2,6-dimethyl-biphenyl-4-yl) ester (8.8 g, 25.8 mmol) inmethanol (65 mL) is added sodium methoxide (1.39 g, 25.8 mmol). Thereaction mixture is heated to reflux overnight. After cooling to roomtemperature, the reaction mixture is neutralized with 5N HCl,concentrated to ⅓ volume, treated with brine, and extracted intodichloromethane. The aqueous layer is back-extracted withdichloromethane, and the combined organic layers are dried andconcentrated. The resulting residue is applied to silica gel and elutedusing hexanes with an ethyl acetate gradient from 0% to 50% to give thetitle compound (5.84 g).

Preparation 11 2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-thiol

The titled compound is prepared in a substantially similar manner asdescribed in Preparation 10 using2,6-dimethyl-4′-(trifluoromethyl)biphenyl-4-ol. MS (ES): 281.1 [M−H]⁻.

Example 1(R,S)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide

Step A.(R,S)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid ethyl ester

A solution of (R,S)-5-(1-hydroxy-2-methyl-propyl)-thiophene-2-carboxylicacid ethyl ester (1.26 g, 5.52 mmol) and4′-tert-butyl-2,6-dimethyl-biphenyl-4-thiol (1.64 g, 6.07 mmol) in1,2-dichloroethane (22 mL) is treated with zinc iodide (1.76 g, 5.52mmol) and stirred overnight at room temperature. The reaction mixture isthen partitioned between water and dichloromethane. The aqueous layer isback-extracted with dichloromethane, and the combined organic layers aredried, filtered, and concentrated. The resulting residue is applied tosilica gel and eluted using hexanes with an ethyl acetate gradient from0% to 40% to give(R,S)-5-[1-(4′-tert-butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid ethyl ester (2.08 g). MS (ES): 481.1 [M+H]⁺.

Step B.(R,S)-5-[1-(4′-tert-butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid

To a mixture of(R,S)-5-[1-(4′-tert-butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid ethyl ester (2.08 g, 4.33 mmol) in ethanol (43 mL) is added sodiumhydroxide (5N aqueous, 4.33 mL) at room temperature, and stirredovernight. The reaction mixture is acidified by 1N HCl (4.42 mL),extracted into ethyl acetate, dried and concentrated, then dried undervacuum, giving(R,S)-5-[1-(4′-tert-butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid (1.82 g). MS (ES): 451.2 [M−H]⁻.

Step C.(R,S)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid cyanomethyl-amide

To a mixture of(R,S)-5-[1-(4′-tert-butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid (342 mg, 0.756 mmol) in DMF (7.6 mL) is addedN-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (290 mg,1.512 mmol), 1-hydroxybenzotriazole hydrate (123 mg, 0.907 mmol), anddiisopropylethylamine (0.264 mL, 1.512 mmol) at room temperature, andstirred 10 min. The mixture is then treated with aminoacetonitrilehydrochloride (84 mg, 0.907 mmol), and stirred overnight. The reactionmixture is treated with 0.1N HCl and extracted into ethyl acetate twice.The combined organic layers are washed with brine, dried andconcentrated, and the resulting residue is applied to silica gel andeluted using hexanes with an ethyl acetate gradient from 0% to 70% togive(R,S)-5-[1-(4′-tert-butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid cyanomethyl-amide (289 mg). MS (ES): 491.1 [M+H]⁺.

Step D.(R,S)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide

A solution of(R,S)-5-[1-(4′-tert-butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid cyanomethyl-amide (195 mg, 0.397 mmol) in toluene (8 mL) is treatedwith triethylamine hydrochloride (275 mg, 2 mmol) and sodium azide (130mg, 2 mmol), then heated at reflux overnight (thermal conditions) oralternatively, placed in a CEM microwave reactor for 20 min. (300 W,180° C., N₂ cooling) (microwave conditions). After cooling to roomtemperature, the reaction mixture is partitioned between ethyl acetateand water. The aqueous layer is back-extracted with ethyl acetate, thecombined organic layers are dried and concentrated, and the resultingresidue is loaded onto C₁₈ and eluted using water with an acetonitrilegradient from 15% to 100% to give the title compound (91 mg). MS (ES):534.2 [M+H]⁺.

Examples 2 to 7 are prepared in a substantially similar manner using thethermal conditions as described in Example 1, Step D.

Example 2(±)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide

MS (ES): 520.3 [M+H]⁺.

Example 3(±)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-butyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide

MS (ES): 534.3 [M+H]⁺.

Example 4(±)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-pentyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide

MS (ES): 548.3 [M+H]⁺.

Example 5(±)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-3-methyl-butyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide

MS (ES): 548.0 [M+H]⁺.

Example 6((±)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-3,3-dimethyl-butyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide

MS (ES): 562.0 [M+H]⁺.

Example 7(±)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide

MS (ES): 548.3 [M+H]⁺.

Examples 8 to 13 are prepared in a substantially similar manner usingthe microwave conditions as described in Example 1, Step D.

Example 8(±)-5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide

MS (ES): 546.0 [M+H]⁺.

Example 9(±)-5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide

MS (ES): 545.8 [M+H]⁺.

Example 10(±)-5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide

MS (ES): 560.0 [M+H]⁺.

Example 11(±)-5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-3-methyl-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide

MS (ES): 560.0 [M+H]⁺.

Example 12(±)-5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-3,3-dimethyl-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide

MS (ES): 574.0 [M+H]⁺.

Example 13(±)-5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-pentyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide

MS (ES): 560.0 [M+H]⁺.

Example 145-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide (Isomer 1)

Step A.5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid cyanomethyl-amide (Isomer 1)

(±)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid cyanomethyl-amide (500 mg) is separated by chiral HPLC (column:Chiralpak AD 4.6×150 mm; eluent: 15/85 3A ethanol/heptane, with 12%dimethylethylamine; flow rate: 0.6 mL/min; UV absorbance wavelength: 300nm) to provide5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid cyanomethyl-amide (Isomer 1) (220 mg).

Step B.5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide (Isomer 1)

A solution of5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid cyanomethyl-amide (Isomer 1) (220 mg, 0.436 mmol) in toluene (8.7mL) is treated with triethylamine hydrochloride (300 mg, 2.18 mmol) andsodium azide (142 mg, 2.18 mmol), then heated at reflux overnight(thermal conditions) or alternatively, placed in a CEM microwave reactorfor 20 min (300 W, 180° C., N₂ cooling) (microwave conditions). Aftercooling to room temperature, the reaction mixture is partitioned betweenethyl acetate and water. The aqueous layer is back-extracted with ethylacetate, and the combined organic layers are dried and concentrated togive the title compound (105 mg). MS (ES): 548.3 [M+H]⁺.

Example 15 to 17 are prepared in a substantially similar manner usingthe thermal conditions as describe in Example 14, Step B.

Example 155-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide (Isomer 2)

MS (ES): 548.3 [M+H]⁺.

Example 165-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide (Isomer 1)

MS (ES): 519.8 [M+H]⁺.

Example 175-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide (Isomer 2)

MS (ES): 520.3 [M+H]⁺.

Examples 18 to 29 are prepared in a substantially similar manner usingthe microwave conditions as described in Example 14, Step B.

Example 185-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 1)

MS (ES): 546.0 [M+H]⁺.

Example 195-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 2)

MS (ES): 546.0 [M+H]⁺.

Example 205-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 1)

MS (ES): 546.0 [M+H]⁺.

Example 215-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 2)

MS (ES): 546.0 [M+H]⁺.

Example 225-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amideb (Isomer 1)

MS (ES): 559.8 [M+H]⁺.

Example 235-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amideb (Isomer 2)

MS (ES): 559.8 [M+H]⁺.

Example 245-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-3-methyl-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 1)

MS (ES): 560.0 [M+H]⁺.

Example 255-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-3-methyl-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 2)

MS (ES): 560.0 [M+H]⁺.

Example 265-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-3,3-dimethyl-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 1)

MS (ES): 573.8 [M+H]⁺.

Example 275-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-3,3-dimethyl-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 2)

MS (ES): 573.8 [M+H]⁺.

Example 285-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-pentyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 1)

MS (ES): 559.8 [M+H]⁺.

Example 295-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-pentyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 2)

MS (ES): 559.8 [M+H]⁺.

The compound of Formula I is preferably formulated in a unit dosage formprior to administration. Therefore, yet another embodiment of thepresent invention is a pharmaceutical composition comprising a compoundof Formula I and one or more pharmaceutically acceptable carriers,diluents or excipients. In such form, the preparation is subdivided intosuitably sized unit doses containing appropriate quantities of theactive components, e.g., an effective amount to achieve the desiredpurpose. Such pharmaceutical compositions and processes for preparingsame are well known in the art. See, e.g., REMINGTON: THE SCIENCE ANDPRACTICE OF PHARMACY (A. Gennaro, et al., eds., 19^(th) ed., MackPublishing Co., 1995). The particular dosage of a compound of formula(1) or a pharmaceutically acceptable salt thereof required to constitutean effective amount according to this invention will depend upon theparticular circumstances of the conditions to be treated. Preferably thecompound is administered orally. The quantity of the inventive activecomposition in a unit dose of preparation may be generally varied oradjusted from about 0.01 milligrams to about 1,000 milligrams,preferably from about 0.01 to about 950 milligrams, more preferably fromabout 0.01 to about 500 milligrams, and typically from about 1 to about250 milligrams, according to the particular application. The actualdosage employed may be varied depending upon the patient's age, sex,weight and severity of the condition being treated. Such techniques arewell known to those skilled in the art. Generally, the human oral dosageform containing the active ingredients can be administered 1 or 2 timesper day. Considerations such as dosage, route of administration, andfrequency of dosing are best decided by the attending physician.

The compositions of the invention may be formulated so as to providequick, sustained or delayed release of the active ingredient afteradministration to the patient. The compositions of the present inventionmay be formulated in sustained release form to provide the ratecontrolled release of any one or more of the components or activeingredients to optimize the therapeutic effects, i.e., glucagon receptorantagonist activity and the like. Suitable dosage forms for sustainedrelease include layered tablets containing layers of varyingdisintegration rates or controlled release polymeric matricesimpregnated with the active components and shaped in tablet form orcapsules containing such impregnated or encapsulated porous polymericmatrices.

There is increasing evidence that glucagon plays an important role inglucose homeostasis. Compounds of Formula I are effective as antagonistsor inverse agonists of the glucagon receptor, and thus inhibit theactivity of the glucagon receptor. More particularly, these compoundsare selective antagonists or inverse agonists of the glucagon receptor.As selective antagonists or inverse agonists, the compounds of Formula Iare useful in the treatment of diseases, disorders, or conditionsresponsive to the inactivation of the glucagon receptor, including butnot limited to diabetic and other glucagon related disorders. It isexpected that selective antagonists or inverse agonists of the glucagonreceptor will lower plasma glucose levels and thus prevent or treatdiabetic and other glucagon related metabolic disorders.

Pharmacological Methods

In the following section binding assays as well as functional assaysuseful for evaluating the efficiency of the compounds of the inventionare described. Binding of compounds to the glucagon receptor may bedetermined in a competition binding assay using the cloned humanglucagon receptor, and selectivity against the hGlp1 receptor.Antagonism may be determined as the ability of the compounds to inhibitthe amount of cAMP formed in the assay in the presence of 5 nM glucagon.

Glucagon Receptor (hGlucR) Binding Assay

The receptor binding assay uses cloned human glucagon receptor (Lok S,Kuijper J L, Jelinek L J, Kramer J M, Whitmore T E, Sprecher C A,Mathewes S, Grant F J, Biggs S H, Rosenberg G B, et al. Gene 140 (2),203-209 (1994)) isolated from 293HEK membranes. The hGlucR cDNA issubcloned into the expression plasmid phD (Trans-activated expression offully gamma-carboxylated recombinant human protein C, an antithromboticfactor. Grinnell, B. W., Berg, D. T., Walls, J. and Yan, S. B.Bio/Technology 5: 1189-1192 (1987)). This plasmid DNA is transfectedinto 293 HEK cells and selected with 200 μg/mL Hygromycin.

Crude plasma membranes are prepared using cells from suspension culture.The cells are lysed on ice in hypotonic buffer containing 25 mM TrisHCL, pH 7.5, 1 mM MgCl2, DNAse1, 20 u/mL, and Roche CompleteInhibitors-without EDTA. The cell suspension is homogenized with a glassdounce homogenizer using a Teflon pestle for 25 strokes. The homogenateis centrifuged at 4 degrees C. at 1800×g for 15 mins. The supernate iscollected and the pellet is resuspended in hypotonic buffer andrehomogenized. The mixture is centrifuged at 1800×g for 15 mins. Thesecond supernate is combined with the first supernate. The combinedsupernates are recentrifuged at 1800×g for 15 mins to clarify. Theclarified supernate is transferred to high speed tubes and centrifugedat 25000×g for 30 minutes at 4 degrees C. The membrane pellet isresuspended in homogenization buffer and stored as frozen aliquots at−80 degree C. freezer until needed.

Glucagon is radioiodinated by I-125-lactoperoxidase procedure andpurified by reversed phase HPLC at Perkin-Elmer/NEN (NEX207). Thespecific activity is 2200 Ci/mmol. Kd determination is performed byhomologous competition instead of saturation binding due to highpropanol content in the I-125 glucagon material. The Kd is estimated tobe 3 nM and is used to calculate Ki values for all compounds tested.

The binding assays are carried out using a Scintillation Proximity Assay(Amersham) with WGA beads previously blocked with 1% fatty acid free BSA(ICN). The binding buffer contains 25 mM Hepes, pH 7.4, 2.5 mM CaCl₂, 1mM MgCl₂, 0.1% fatty acid free BSA, (ICN), 0.003% tween-20, and RocheComplete Inhibitors without EDTA. Glucagon is dissolved in 0.01 N HCl at1 mg/mL and immediately frozen at −80 degrees C. in 30 μl aliquots. Theglucagon aliquot is diluted and used in binding assays within an hour.Test compounds are dissolved in DMSO and serially diluted in DMSO. 10 uldiluted compounds or DMSO is transferred into Corning 3632, opaque clearbottom assay plates containing 90 μl assay binding buffer or coldglucagon (NSB at 1 μM final). 50 μl of 1-125 glucagon (0.15 nM final inreaction), 50 μl of membranes (300 μg/well), and 40 μl of WGA beads (150mgs/well) are added, covered, and mixed end over end. Plates are readwith a MicroBeta after 14 hours of settling time at room temp.

Results are calculated as a percent of specific I-125-glucagon bindingin the presence of compound. The absolute EC50 dose of compound isderived by non-linear regression of percent specific binding ofI-125-glucagon vs. the dose of compound added. The EC50 dose isconverted to Ki using the Cheng-Prusoff equation (Cheng Y., Prusoff W.H., Biochem. Pharmacol. 22, 3099-3108, 1973).

Glucagon-Like-Peptide 1 (Glp1-R) Receptor Binding Assay

The receptor binding assay uses cloned human glucagon-like peptide 1receptor (hGlp1-R) (Graziano M P, Hey P J, Borkowski D, Chicchi G G,Strader C D, Biochem Biophys Res Commun. 1993 Oct. 15; 196(1):141-6)isolated from 293HEK membranes. The hGlp1-R cDNA is subcloned into theexpression plasmid phD (Trans-activated expression of fullygamma-carboxylated recombinant human protein C, an antithromboticfactor. Grinnell, B. W., Berg, D. T., Walls, J. and Yan, S. B.Bio/Technology 5: 1189-1192 (1987)). This plasmid DNA is transfectedinto 293 HEK cells and selected with 200 μg/mL Hygromycin.

Crude plasma membrane is prepared using cells from suspension culture.The cells are lysed on ice in hypotonic buffer containing 25 mM TrisHCl, pH 7.5, 1 mM MgCl₂, DNAse, 20 W/mL, and Roche Complete Inhibitorswithout EDTA. The cell suspension is homogenized with a glass douncehomogenizer using a Teflon pestle for 25 strokes. The homogenate iscentrifuged at 4 degrees C. at 1800×g for 15 mins. The supernate iscollected and the pellet is resuspended in hypotonic buffer andrehomogenized. The mixture is centrifuged at 1800×g for 15 mins. Thesecond supernate is combined with the first supernate. The combinedsupernates are recentrifuged at 1800×g for 15 mins to clarify. Theclarified supernate is transferred to high speed tubes and centrifugedat 25000×g for 30 minutes at 4 degrees C. The membrane pellet isresuspended in homogenization buffer and stored as frozen aliquots in−80 degree C. freezer until use.

Glucagaon-like peptide 1 (Glp-1) is radioiodinated by theI-125-lactoperoxidase procedure and purified by reversed phase HPLC atPerkin-Elmer/NEN (NEX308). The specific activity is 2200 Ci/mmol. Kddetermination is performed by homologous competition instead ofsaturation binding due to high propanol content in the 1-125 Glp-1material. The Kd is estimated to be 3 nM and is used to calculate Kivalues for all compounds tested.

The binding assays are carried out using a Scintillation Proximity Assay(Amersham) with wheat germ agglutinin (WGA) beads previously blockedwith 1% fatty acid free BSA (ICN). The binding buffer contains 25 mMHepes, pH 7.4, 2.5 mM CaCl₂, 1 mM MgCl₂, 0.1% fatty acid free BSA,(ICN), 0.003% tween-20, and Roche Complete Inhibitors without EDTA.Glucagon-like peptide 1 is dissolved in PBS at 1 mg/mL and immediatelyfrozen at −80 degrees C. in 30 ul aliquots. The glucagon-like peptidealiquot is diluted and used in binding assays within an hour. Testcompounds are dissolved in DMSO and serially diluted in DMSO. 10 μldiluted compounds or DMSO is transferred into Corning 3632, opaque clearbottom assay plates containing 90 μl assay binding buffer or coldglucagon-like peptide 1 (NSB at 1 μM final). 50 μl of 1-125glucagon-like peptide 1 (0.15 nM final in reaction), 50 μl of membranes(600 μg/well), and 40 μl of WGA beads (150 μgs/well) are added, covered,and mixed end over end. Plates are read with a MicroBeta after 14 hoursof settling time at room temp.

Results are calculated as a percent of specific I-125-glucagon-likepeptide 1 binding in the presence of compound. The absolute EC50 dose ofcompound is derived by non-linear regression of percent specific bindingof I-125-glucagon-like peptide 1 vs. the dose of compound added. TheEC50 dose is converted to Ki using the Cheng-Prusoff equation (Cheng Y.,Prusoff W. H., Biochem. Pharmacol. 22, 3099-3108, 1973).

Glucagon-Stimulated cAMP Functional Antagonist Assay

The cAMP functional assay uses the same cloned human glucagon receptorcell line isolated for the hGlucR binding assay described above. Cellsare stimulated with a mixture of an EC80 dose of glucagon in thepresence of compound. The cAMP generated within the cell is quantitatedusing an Amplified Luminescent Proximity Homogeneous Assay, AlphaScreen, from Perkin Elmer (6760625R).

Briefly, cAMP within the cell competes for binding of biotinylated cAMPfrom the kit to a coated anti-cAMP antibody Acceptor bead and astrepavidin coated Donor bead. As the cAMP level within the cellincreases, a disruption of the Acceptor bead-biotinlyated cAMP—Donorbead complex occurs and decreases the signal.

Glucagon is dissolved in 0.01 N HCl at 1 mg/mL and immediately frozen at−80 degrees C. in 30 ul aliquots. The glucagon aliquot is diluted andused in the functional assay within an hour. Cells are harvested fromsub-confluent tissue culture dishes with Enzyme-Free Cell DissociationSolution, (Specialty Media 5-004-B). The cells are pelleted at low speedand washed 3 times with assay buffer [25 mM Hepes in HBSS-with Mg and Ca(GIBCO, 14025-092) with 0.1% Fatty Acid Free BSA (ICN)] then diluted toa final concentration of 250,000 cells per mL. Compounds are seriallydiluted into DMSO then diluted into assay buffer with a 3× concentrationof glucagon and 3% DMSO. The EC80 of glucagon is pre-determined from afull glucagon dose response and represents the dose at which glucagonsproduces an 80% of the maximal glucagon response. A mixture ofbiotinylated cAMP (1 unit/well final) from the Alpha Screen Kit and3×IBMX (1500 μM) is prepared in Assay Buffer.

The functional assay is performed in 96 well, low-volume, white,poylstyrene Costar Plates (3688). The biotinylated cAMP/IBMX mixture,0.02 mLs, is placed into each well, followed by addition of 0.02 mLs ofglucagon dose response, cAMP standard curve, or compound/glucagonmixtures. The reaction is started by addition of 0.02 mLs of cells(5000/well final). After 60 minutes at room temperature, the reaction isstopped by the addition of 0.03 mLs of Lysis Buffer [10 mM Hepes, pH7.4, 1% NP40, and 0.01% fatty acid free BSA (ICN) containing 1 uniteach/well of Acceptor and Donor beads from the Alpha Screen Kit]. LysisBuffer addition is performed under a green light to prevent bleaching ofthe detection beads. The plates are wrapped in foil and left toequilibrate overnight at room temperature. The plates are read on aPackard Fusion™-α Instrument.

Alpha screen units are converted to pmoles cAMP generated per well basedupon the cAMP standard curve. The pmoles cAMP produced in the presenceof compound are converted to % of a maximal response with the EC80 doseof glucagon alone. With each experiment, the dose of glucagon needed toproduce a 50% response of pmoles cAMP is determined. This EC50 dose isused to normalize results to a Kb using a modified Cheng-Prusoffequation (Cheng Y., Prusoff W. H., Biochem. Pharmacol. 22, 3099-3108,1973), where Kb=(EC50 compound)/[1+(pM glucagon used/EC50 in pM forglucagon dose response)].

The compounds according to the invention preferably have a Ki value ofno greater than 50 μM as determined by the Glucagon Receptor (hGlucR)Binding Assay disclosed herein. More preferably, the compounds accordingto the invention have a Ki value of less than 5 μM, preferably of lessthan 500 nM and even more preferred of less than 100 nM as determined bythe Glucagon Receptor (hGlucR) Binding Assay disclosed herein.Generally, the compounds according to the invention show a higheraffinity for the glucagon receptor compared to the GLP-1 receptor, andpreferably have a higher binding affinity to the glucagon receptor thanto the GLP-1 receptor. All of the examples provided herein have a Kivalue of less than 10 μM.

The results are given below for the indicated compound.

TABLE 1 Example Ki (nM)

18.4

36.0

From the above description, one skilled in the art can ascertain theessential characteristics of the present invention, and withoutdeparting from the spirit and scope thereof, can make various changesand modifications of the invention to adapt it to various usages andconditions. Thus, other embodiments are also within the claims.

1. A compound structurally represented by Formula I

or a pharmaceutically acceptable salt thereof wherein: R1 and R2 areindependently —H or -halogen; R3 is —(C₁-C₈)alkyl (optionallysubstituted with 1 to 3 halogens), —(C₃-C₇)cycloalkyl,—(C₁-C₆)alkyl-(C₃-C₇)cycloalkyl, or —(C₃-C₇)cycloalkyl-(C₁-C₆)alkyl(optionally substituted with 1 to 3 halogens); R4 and R5 areindependently —H, -halogen, -hydroxy, hydroxymethyl, —CN,—(C₁-C₇)alkoxy, —(C₂-C₇)alkenyl, or —(C₁-C₆)alkyl (optionallysubstituted with 1 to 3 halogens); R6 is

wherein the zig-zag mark shows the point of attachment to the parentmolecule; R7 and R8 are independently —H, -halogen, —(C₁-C₆)allyl(optionally substituted with 1 to 3 halogens), —(C₁-C₆)alkoxy,—(C₃-C₇)cycloalkyl, —C(O)R10, —COOR10, —OC(O)R10, —OS(O)₂R10, —SR10,—S(O)R10, —S(O)₂R10, or —O(C₂-C₇)alkenyl; R9 is independently —H,-halogen, —CN, —(C₃-C₇)cycloalkyl, —C(O)R10, —COOR10, —OC(O)R10,—OS(O)₂R10, —SR10, —S(O)R10, —S(O)₂R10, or —O(C₂-C₇)alkenyl,—(C₁-C₃)alkoxy (optionally substituted with 1 to 3 halogens), or—(C₁-C₆)alkyl (optionally substituted with 1 to 3 halogens); and R10 isindependently at each occurrence -hydrogen, or —(C₁-C₆)alkyl (optionallysubstituted with 1 to 3 halogens).
 2. A compound of claim 1, or apharmaceutically acceptable salt thereof, wherein R1 and R2 are —H; R3is —(C₁-C₈)alkyl (optionally substituted with 1 to 3 halogens),—(C₃-C₆)cycloalkyl, —(C₁-C₆)alkyl-(C₃-C₆)cycloalkyl, or—(C₃-C₆)cycloalkyl-(C₃-C₆)alkyl (optionally substituted with 1 to 3halogens); R4 and R5 are independently —H, -halogen, or —(C₁-C₆)alkyl(optionally substituted with 1 to 3 halogens); R6 is

wherein the zig-zag mark shows the point of attachment to the parentmolecule; R7 and R8 are independently —H, -halogen, —(C₁-C₃)allyl(optionally substituted with 1 to 3 halogens), or —(C₁-C₃)alkoxy; and R9is independently —H, -halogen, or —(C₁-C₆) alkyl (optionally substitutedwith 1 to 3 halogens).
 3. A compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein R1 and R2 are —H; R3 is —(C₁-C₈)alkyl(optionally substituted with 1 to 3 halogens), —(C₃-C₆)cycloalkyl,—(C₁-C₆)alkyl-(C₃-C₆)cycloalkyl, or —(C₃-C₆)cycloalkyl-(C₁-C₆)alkyl(optionally substituted with 1 to 3 halogens); R4 and R5 areindependently —H, -halogen, or —CH₃ (optionally substituted with 1 to 3halogens); R6 is

wherein the zig-zag mark shows the point of attachment to the parentmolecule; R7 and R8 are independently —H, or -halogen; and R9 isindependently —(C₁-C₆)alkyl (optionally substituted with 1 to 3halogens).
 4. The compound of claim 1, or a pharmaceutically acceptablesalt thereof, wherein R1 and R2 are —H; R3 is —(C₁-C₈)allyl (optionallysubstituted with 1 to 3 halogens), —(C₃-C₆)cycloalkyl,—(C₁-C₆)alkyl-(C₃-C₆)cycloalkyl, or —(C₃-C₆)cycloalkyl-(C₁-C₆)alkyl(optionally substituted with 1 to 3 halogens); R4 and R5 are —CH₃(optionally substituted with 1 to 3 halogens) and each occupies aposition adjacent to R6 on the phenyl ring to which R6 is attached; R6is

wherein the zig-zag mark shows the point of attachment to the parentmolecule; R7 and R8 are —H; and R9 is independently —(C₁-C₆) alkyl(optionally substituted with 1 to 3 halogens).
 5. The compound of claim1, or a pharmaceutically acceptable salt thereof, wherein R1 and R2 areindependently hydrogen or halogen; R3 is methyl, ethyl, propyl,isopropyl, butyl, pentyl, hexyl, heptyl, octyl, 3,3-dimethylbutyl,2-methylpropyl, 3-methyl-butyl, tertbutyl, 4-methylpentyl,2,2-dimethylpropyl, 3,3,3-trifluoropropyl, 4,4,4-trifluorobutyl,cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl; R4 and R5 areindependently hydrogen, methyl, ethyl, tertbutyl, cyclohexyl, pentyl,isopropoxy, chloro, fluoro, bromo, hydroxy, trifluoromethyl, —CN,methoxy, hydroxymethyl, 4-methylpentyloxy, or pentyloxy; R7 and R8 areindependently hydrogen, fluoro, chloro, methyl, ethyl, pentyl,isopropyl, tertbutyl, trifluoromethyl, acetyl, 2-methylpropyl, methoxy,cyclohexyl, or trifluoromethoxy; and R9 is hydrogen, bromo, fluoro,methyl, tertbutyl, trifluoromethyl, or isopropyl.
 6. The compound ofclaim 1, selected from the group consisting of formulae XI to XII;Formula Number Structure X1

X2

X3

X4

X5

X6

X7

X8

X9

X10

X11

or a pharmaceutically acceptable salt thereof.
 7. A compound of claim 1selected from the group consisting of(R,S)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide;(±)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide;(±)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-butyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide;(±)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-pentyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide;(±)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-3-methyl-butyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide;((±)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-3,3-dimethyl-butyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide;(±)-5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide;(±)-5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide;(±)-5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide;(±)-5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide;(±)-5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-3-methyl-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide;(±)-5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-3,3-dimethyl-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide; (±)-5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-pentyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide;5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide (Isomer 1);5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide (Isomer 2);5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide (Isomer 1);5-[1-(4′-tert-Butyl-2,6-dimethyl-biphenyl-4-ylsulfanyl)-propyl]-thiophene-2-carboxylicacid (2H-tetrazol-5-ylmethyl)-amide (Isomer 2);5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 1);5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 2);5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 1);5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-2-methyl-propyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 2);5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amideb (Isomer 1);5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenylsulfanyl)-2,2-dimethyl-propyl]-thiophene-2-carboxylic acid(1H-tetrazol-5-ylmethyl)-amideb (Isomer 2);5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-3-methyl-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 1);5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-3-methyl-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 2);5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-3,3-dimethyl-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 1);5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-3,3-dimethyl-butyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 2);5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-pentyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 1); and5-[1-(2,6-Dimethyl-4′-trifluoromethyl-biphenyl-4-ylsulfanyl)-pentyl]-thiophene-2-carboxylicacid (1H-tetrazol-5-ylmethyl)-amide (Isomer 2); or a pharmaceuticallyacceptable salt thereof.
 8. A pharmaceutical composition which comprisesa compound or salt of claim 1 and a pharmaceutically acceptable carrier.